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基本情報
登録情報 | データベース: PDB / ID: 7q4d | ||||||
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タイトル | Local refinement structure of the two interacting N-domains of full-length, dimeric, soluble somatic angiotensin I-converting enzyme | ||||||
![]() | Angiotensin-converting enzyme![]() | ||||||
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機能・相同性 | ![]() mononuclear cell proliferation / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() 類似検索 - 分子機能 | ||||||
生物種 | ![]() ![]() | ||||||
手法 | ![]() ![]() ![]() | ||||||
![]() | Lubbe, L. / Sewell, B.T. / Sturrock, E.D. | ||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Cryo-EM reveals mechanisms of angiotensin I-converting enzyme allostery and dimerization. 著者: Lizelle Lubbe / Bryan Trevor Sewell / Jeremy D Woodward / Edward D Sturrock / ![]() 要旨: Hypertension (high blood pressure) is a major risk factor for cardiovascular disease, which is the leading cause of death worldwide. The somatic isoform of angiotensin I-converting enzyme (sACE) ...Hypertension (high blood pressure) is a major risk factor for cardiovascular disease, which is the leading cause of death worldwide. The somatic isoform of angiotensin I-converting enzyme (sACE) plays a critical role in blood pressure regulation, and ACE inhibitors are thus widely used to treat hypertension and cardiovascular disease. Our current understanding of sACE structure, dynamics, function, and inhibition has been limited because truncated, minimally glycosylated forms of sACE are typically used for X-ray crystallography and molecular dynamics simulations. Here, we report the first cryo-EM structures of full-length, glycosylated, soluble sACE (sACE ). Both monomeric and dimeric forms of the highly flexible apo enzyme were reconstructed from a single dataset. The N- and C-terminal domains of monomeric sACE were resolved at 3.7 and 4.1 Å, respectively, while the interacting N-terminal domains responsible for dimer formation were resolved at 3.8 Å. Mechanisms are proposed for intradomain hinging, cooperativity, and homodimerization. Furthermore, the observation that both domains were in the open conformation has implications for the design of sACE modulators. | ||||||
履歴 |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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-検証レポート
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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-関連構造データ
関連構造データ | ![]() 13803MC ![]() 7q3yC ![]() 7q49C ![]() 7q4cC ![]() 7q4eC C: 同じ文献を引用 ( M: このデータのモデリングに利用したマップデータ |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
電子顕微鏡画像生データ | ![]() Data size: 3.8 TB Data #1: Unaligned multi-frame cryo-EM micrographs of human somatic angiotensin I-converting enzyme in the apo state [micrographs - multiframe]) |
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リンク
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集合体
登録構造単位 | ![]()
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要素
-タンパク質 / 非ポリマー , 2種, 4分子 AB![](data/chem/img/ZN.gif)
![](data/chem/img/ZN.gif)
#1: タンパク質 | ![]() 分子量: 139614.000 Da / 分子数: 2 / 変異: P576L / 由来タイプ: 組換発現 詳細: Soluble secreted form of human somatic angiotensin I-converting enzyme terminating at Ser1211 由来: (組換発現) ![]() ![]() 発現宿主: ![]() ![]() ![]() 参照: UniProt: P12821, ![]() ![]() #7: 化合物 | |
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-糖 , 6種, 16分子 ![](data/chem/img/NAG.gif)
#2: 多糖 | ![]() #3: 多糖 | beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta- ...beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose ![]() #4: 多糖 | ![]() #5: 多糖 | ![]() #6: 多糖 | ![]() #8: 糖 | ![]() |
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-詳細
研究の焦点であるリガンドがあるか | Y |
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-実験情報
-実験
実験 | 手法: ![]() |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: ![]() |
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試料調製
構成要素 | 名称: Full-length, soluble, dimeric somatic angiotensin I-converting enzyme タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT | ||||||||||||||||||||
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分子量 | 値: 0.279 MDa / 実験値: NO | ||||||||||||||||||||
由来(天然) | 生物種: ![]() ![]() | ||||||||||||||||||||
由来(組換発現) | 生物種: ![]() ![]() ![]() プラスミド ![]() | ||||||||||||||||||||
緩衝液 | pH: 7.5 / 詳細: Solutions were prepared with deionized water | ||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 1.5 mg/ml / 包埋: NO / シャドウイング: NO / 染色![]() ![]() 詳細: The protein was stored at 3.0mg/ml in 50mM HEPES (pH 7.5) and diluted immediately prior to grid preparation | ||||||||||||||||||||
試料支持 | グリッドの材料: COPPER / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: Quantifoil R2/2 | ||||||||||||||||||||
急速凍結![]() | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K 詳細: Diluted protein (in buffer containing zinc chloride and sodium chloride) was incubated on ice for 30 minutes after which it was applied to the grid, incubated for 30 seconds, and blotted for ...詳細: Diluted protein (in buffer containing zinc chloride and sodium chloride) was incubated on ice for 30 minutes after which it was applied to the grid, incubated for 30 seconds, and blotted for 3 seconds before plunging |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源![]() ![]() |
電子レンズ | モード: BRIGHT FIELD![]() ![]() |
試料ホルダ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 3 sec. / 電子線照射量: 43 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 1 / 実像数: 11628 詳細: Images were recorded in super-resolution mode with 40 frames per image |
画像スキャン | 横: 5760 / 縦: 4092 |
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解析
ソフトウェア |
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EMソフトウェア |
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CTF補正![]() | 詳細: cryoSPARC patch-based CTF estimation / タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 1727045 詳細: Topaz-denoising was performed on all curated micrographs, a Topaz model trained on this dataset, and used for picking | ||||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 点対称性![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||
3次元再構成![]() | 解像度: 3.78 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 215632 詳細: Local refinement (C1) with a mask around the two interacting N-domains following C-domain particle subtraction in RELION and C2 symmetry expansion in cryoSPARC クラス平均像の数: 1 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL / Target criteria: Correlation coefficient | ||||||||||||||||||||||||||||||||||||||||||||||||||
精密化 | 交差検証法: NONE 立体化学のターゲット値: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||||||||||||||||||||||||||||
原子変位パラメータ | Biso mean: 81.84 Å2 | ||||||||||||||||||||||||||||||||||||||||||||||||||
拘束条件 |
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